Instrumental Methods of Analysis 1. Introduction 1.Writing Intensive 1.Rewrites are Required 2.Large Amount of Excel Data Management 2.Work 1.Individual 1.Work Log 2.Own Sample 3.Exams 4.Lab Book pages 2.Group 1.Division and Attribution of Labor (Accountability) 2.Reports 3.Calibration Curves 3.Single Analyte (Skill Development and Cross Comparison Methods) 1.Overlapping Themes 1.Statistics/Sampling 2.Solution Chemistry 3.EDTA 4.Isotopes 5.Phase Separations

1.Writing Intensive 2.Work 3.Single Analyte 4.Problem Based What is best method for lead analysis? What is best method for This particular sample? 5.Ethics and Service Learning 1.How do we know? 2.Critical Thinking Instrumental Methods of Analysis 1. Introduction

Electronics, Spectroscopy, Electrochemistry, Chromatography Confusion “Too much from instructor” information Iodide Electrolysis Chromium & permanganate UV-Vis Fluoride Ion Selective Electrode Quinine Fluorescence Cu Graphite Furnace Atomic Absorption DMSO Infrared Spectroscopy Methanol Gas Chromatography 7 different analytes 7 different methods How do we select Or compare methods/ Instruments?

Figures of Merit ThemeKnowledge Pb Anodic Stripping Voltammetry Pb Dithizone UV-Vis Tetraethyl 207 Pb MS Pb Ion Selective Electrode Pb EDTA Infrared Spectroscopy EDTA Pb HPLC 207 Pb in EDTA by proton NMR Pb Atomic Absorption Calcein Blue Pb Fluorescence Quenching Choose Method: Cross Compare/measure LOD Role of solution pH important Pb Limit of detection? Pb Linear range? Pb Selectivity? Pb Signal/Noise?

WeekTopic“Lab”Concept Stat Chem Isotope Ethics 1 Pb & Society; Statistics and Ethics Pototo populations x x 2 Signals & Noise; LOD Analog & Digital filters x 3 Relevant Chemistry; Exam I ISE & Pb(OH) 2 x 4 Intro Spectroscopy IR & UV-Vis x 5 Molecular Spectroscopy IR & UV-Vis x 6 Fluorescence and AA Fluorescence and FAA x x 7 AA and Vibrational, Exam II Fluorescence and FAA x x 8 Break, NMR Break 9 NMR, Off campus sampling NMR, ASV x x x 10 MS, NMR, ASV x x x 11 Intro Electrochem., Select Method sample prep x x x 12 Potentiometry, sample analysis x x 13 ASV, Take home exam III ICP-MS, GC x x 14 Separations ICP-MS, GC x x 15 Separations How do we know? Ethics x x

Why Lead? Health

Same size, Shape, chemistry Different Orbitals, Different total Number of electrons

Warren et al, TIBS, 1998, 23, 217

Current measurements no longer rely on zinc protoporphyrin as PbB effects on fetal development can be detected at values of PbB <5 ug/dL

ALAD dehydratase with lead binding site An estimated 40% of lead in blood plasma bound to ALAD Wetmur, 1994

Enhanced ALA concentration leads to tautomers which easily form radicals which may attack the cell wall - leading to early cell death. Heme dies about 20% earlier Hammond and Dietrich, 1990

Because lead affected ALAD resulting in overproduction of ALA ALA may also affect the function of GABA but a close similarity in molecular structure GABA controls leaf tip growth, and brain development. Function changes with time, so growing tips and infants differentially affected from adults

Herlenius and Lagercrantz, 2001 Ben-Ari1, 2001 Certain portions of the brain more greatly affected: prefrontal cortex: problem solving hippocampus (memory) cerebellum (motor coordination, body movement, posture and balance)

Why Lead? Everywhere

Lead as an ore in the earth relatively benign Human activity distributes lead into the environment

Modern major uses 1. Pigments 2. Leaded Gasoline 3. Munitions

Eliminating Childhood Lead Poisoning: A Federal Strategy Targeting Lead Pain Hazards, Feb. 2000

Cement block ash Sewage sludge landfill incinerate Mine reclaim fertilizer groundwater Deposit locally Bottom sludge Deposit elsewhere Fishing sinkers & shot Plant uptake Flow to groundwater To air Other Ecosytem Impacts of Lead

2009 Class Project

Loyola Chem Sr. S Growing Scientists 20 5th graders map park soil lead F 1994Northwest Incinerator Austin homes: Yard Soil Lead F 1995 Northwest Incinerator Grid map House Dust Uptown 5th graders sample and tour labs F 1996F 1997 House Dust Little village Boys and Girls Club (failed) F 1998Northwest Incinerator Tight grid and isotopes F Bethel New Life: Brownfield Soil sampling F 2000 Orozco School: House Dust F 2001Pullman: White Lead Factory House Dust West Pullman White Lead factory F 2002 Alanah Fitch S 2008 Lead in Toys Evanston Childcare National Award for The Class ACS Mtg poster

Homes Municipal Waste Incinerator Incinerator 1994, 1995, 1998 Lead Elimination Action Drive West-side Alliance for a Safe Environment (WASTE) Center for Neighborhood Technology Austin Neighborhood Council Pb?

Wind Background Soil lead “Shadow” depends on Stack height Largest deposition Is here Decreasing amts Incinerator 1994, 1995, 1998

Students meet with elderly ladies who let them sample soils. The ladies discuss their concerns. sampling Control

4915 W. Kamerling W. Potomac W. Crystal garage Incinerator 1994, 1995, 1998 Front Back Soil Lead in parts per million (ppm)

BRICK N. Ashland 1538 FRAME 6729 N. Ashland Incinerator 1994, 1995, 1998 Comparison homes, 17 mile away

Second year: Resample with respect to the wind; also allowing for up and down wind comparisons. Wins a National Award. Luke later goes to Croatia On UN project on Lead dispersion by gasoline Incinerator 1994, 1995, 1998

North Chicago Washington Demolished lot Wind Pattern Cicero Pulaski 644 m Orr High School Incinerator 1994, 1995, 1998 This data presented To Mayor’s advisory council Class final: enter Busch National Env. Award competition – Place third

< 0.5 mile from incinerator > 0.5 mile from incinerator Incinerator 1994, 1995, 1998

Wind Background Soil lead “Shadow” depends on Stack height Largest deposition Is here Decreasing amts Incinerator 1994, 1995, 1998

N NNE NE ENE E SE Incinerator 1994, 1995, 1998 Last time: we finally set up an experiment worthy of an An air pollution expert

Across 1the system of pipes and other apparatus for conveying water, liquid wastes, etc., as in a building 3.the fluid that circulates and carries oxygen throughout the human body 4. being placed at risk 8 the quality of degree of being toxic or harmful to plants or animal life 9. the introduction of harmful substances or products into the environment 10.a mixture of pigment and liquid 12. food, eating healthy 14. a pain located in the head 15. the age group affected most by lead poisoning 17. President on the $20 bill 18.a transparent, odorless, tasteless liquid, a compound of hydrogen and oxygen, H20, that constitutes rain, oceans, lakes, rivers, etc to take ill through eating Down 2. a heavy, comparatively soft, malleable, bluish ‑ gray metal. A toxic metal, that has been widely used in paints, plumbing and fixtures, and in some water supply service lines 3. composer thought to have had lead poisoning 5. the portion of the earth's surface consisting of dirt 6. the air, water, minerals, organisms, and all other external factors surrounding and affecting a given organism at any time 7. dry particle of earth 10.a table illustrating the periodic system, in which the chemical elements are shown in related groups 11.. the process of preparing the samples by breaking down the baby wipes into a substance suitable testing 13. a silver ‑ white divalent metal found in limestone, chalk, milk 16. the science that deals with the composition and properties of substances and various elementary forms of matter

2009 – Brainstorm on Projects Short list by next week

Health Effects of Lead

Inhalation depends upon the particle size. This data is for humans

Dissolution of lead carbonate particles in simulated gastric juices as a function of particle size For birds, the uptake will depend upon the pH and grinding action in the gizzard. For ruminants, the uptake will depend upon the lifetime in the digestive organs

94-99% of lead attached to external surfaces of erythrocytes 1-6% lead in plasma, of which 99% attached to proteins ~0.1 to 0.6 % of lead is as the free cation

Ca Ba Active uptake of calcium, apparently to lesser extent other divalent similar sized cations, including lead. Recognition will be similar for various species Calcium uptake is controlled by Vitamin D and growth regulators parathyroid hormone (PTH)

Calcium is stored to the endoplasmic reticulum, lead will also go there

Basophilic Stippling of Dog Blood caused by nucleation of ribonucleoproteins in presence of lead High lead causes deformation of blood cells and shorter lifetime of the red blood cell. Double whammy - production is also affected

Current measurements no longer rely on zinc protoporphyrin as PbB effects on fetal development can be detected at values of PbB <5 ug/dL

Renal biopsy of a 28 year old shipwrecker. Dense intranuclear inclusion body is characterized by a fibrillary outer margin; Cramer, Goyer et al. Brit. J. Indust. Med., hour after exposure lead is found within the kidneys, because kidneys are monitoring blood calcium levels via the glomerular filtration rate

Renal biopsy of a 28 year old shipwrecker. Interstitial fibrosis with atrophy of some tubular lining cells; Cramer, Goyer et al. Brit. J. Indust. Med., 1984

Damage to kidney results in hypertension gout

Thomas Rowlandson’s The King’s Bath

Cruikshank’s gout

Calcium is closely monitored by kidney because it plays a large role in various signalling processes

Calcium bound to calmodulin C. Yang, G. S. Jas, K. Kuczera, J. Biomolecular Structural Dynamics, 2001, 19,

Lead double whammy 1. Low lead turns on triggers 2. Hi lead turns off triggers Calcium serves as a trigger for muscle contractions: troponin C

Colic is a form of muscle spasming So are the “shakes”

Lead, like calcium, is deposited to the hard tissues Implications for the Trophic level of lead in the environment Osteocalcin (bone Gla protein (BGP)) controls calcium deposition and is affected by lead: weaker bones for affected individuals

Longitudinal Section & Cross Section of Bone Trabecular Bone Compact Bone

Seizures can result from lead due to Ca triggers of the synapses

Other Irrelevant Information that Is mildly amusing/interesting on Lead

886 o C -0.2 o C (Au) 230 o C (Ag) Purity of Pb: 99.9% achieved ~5,000 years ago Cuppellation Begins